Dear all,
Two variables:
- Expensiveness
- Time consuming
Using either iTRAQ (protein) or RT-qPCR (RNA) for quantitative (with high sensitivity) expression analysis of a group of 5-15 genes
I appreciate beforehand your kind answers
Best regards
Both methods are complementary. iTraq will give you information about the amount of protein, qPCR with give you data about the amount of mRNA.
In between is the efficiency of translation, stability of mRNA and stability of protein and possible posttranslational modifications, so the data will not necessarily correlate. If you have the machines available and established methods, qPCR is probably easier and cheaper, but I'd recommend defining your analytical needs first depending on the questions you want to answer.
Hello Adrian Muñoz-Bucio
If you need good and quality results and if you have enough funding, I recommend you carry out iTRAQ, and the proteomic results obtained by iTRAQ can be validated using the RT-qPCR and Western blot analysis. In this way you can verify the accuracy of iTRAQ.
Method validation though appears to be a tedious and time-consuming activity, it needs to be done. It provides information which enables the comparability of results from samples analyzed using different methods. Method validation is an essential part of the process of ensuring that measurement results reported are correct and will not let you down under any circumstance.
Good Luck!
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